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为建立一种能检测出临床上绝大多数肠道病毒（ＥＶ）感染的快速、敏感、特异的逆转录聚合酶链反应（ＲＴ－ＰＣＲ）方法，通过对ＥＶ基因组的序列分析，在其５′端非编码区设计一对引物，用该引物建立的ＲＴ－ＰＣＲ方法对３１种ＥＶ标准毒株和３４例无菌性脑膜炎及１１例无菌性脑炎患儿的脑脊液（ＣＳＦ）分别进行扩增，其产物做琼脂糖凝胶电泳及斑点杂交检测。研究表明，３１种ＥＶ标准毒株斑点杂交的敏感度达１０－２～１０－３５０％组织培养感染量（ＴＣＩＤ５０）。３４例无菌性脑膜炎中２１例阳性（６１．８％），１１例无菌性脑炎中８例阳性（７２．７％）；阳性患儿恢复期ＣＳＦ，２例无菌性脑膜炎和１例无菌性脑炎仍为阳性。提示用ＲＴ－ＰＣＲ方法检测ＥＶ感染是值得推广的。
In order to establish a rapid, sensitive and specific reverse transcription-polymerase chain reaction (RT-PCR) method which can detect most clinical EV infections, ’End non-coding region. A pair of primers was designed. The RT-PCR method was used to detect the CSF of 31 EV standard strains and 34 cases of aseptic meningitis and 11 cases of aseptic encephalitis For amplification, the product of agarose gel electrophoresis and dot blot hybridization test. Studies have shown that the sensitivity of 31 EV standard strains spot hybridization 10-2 ~ 10-350% tissue culture infection (TCID50). Among the 34 cases with aseptic meningitis, 21 cases were positive (61.8%) and 11 cases were aseptic encephalitis (72.7%). In the positive children, convalescent CSF and 2 cases of aseptic meningitis And 1 case of aseptic encephalitis is still positive. Prompt RT-PCR method to detect EV infection is worth promoting.